Illuminating tissue samples in wide field fluorescence lifetime imaging microscopy

Abstract

A method of evenly illuminating a tissue sample in a wide field Fluorescence Lifetime Imaging Microscopy (FLIM) and a system for practice thereof. Particularly provided here is a method of evenly illuminating a slide containing a tissue sample in a wide field light source by (i) illuminating the slide containing the tissue sample from more than one side, or (ii) illuminating the slide containing the tissue sample using a square wave excitation, or (iii) illuminating the slide containing the tissue sample using a combination of (i) and (ii) and capturing an individual wide field image or a plurality of wide field images of the tissue sample for accurate medical diagnosis of a medical condition.

What is claimed is: 
     
         1 . A method of evenly illuminating a tissue sample within a wide field Fluorescence Lifetime Imaging Microscopy (FLIM) device, comprising:
 positioning a slide containing the tissue sample within the wide field FLIM device such that a focal point of an objective lens of the wide field FLIM device is within the tissue sample contained within the slide;   illuminating a top surface of the slide with an excitation light source;   illuminating a bottom surface of the slide with the excitation light source; and   capturing an image of a portion of the tissue sample from the top surface of the slide and an image of a portion of the tissue sample from the bottom surface of the slide for diagnosis.   
     
     
         2 . A method of evenly illuminating a tissue sample within a wide Fluorescence Lifetime Imaging Microscopy (FLIM) device, comprising:
 positioning a slide containing the tissue sample within the wide field FLIM device such that a focal point of an objective lens of the wide field FLIM device is within the tissue sample contained within the slide,   illuminating either a top surface of the slide with an excitation light source emitting a square wave or a bottom surface of the slide with the excitation light source emitting the square wave; and   capturing an image of a portion of the tissue sample from the top surface of the slide or an image of a portion of the tissue sample from the bottom surface of the slide for diagnosis.